Chi Mei Medical Center, Tainan 71004, Taiwan.
National Health Research Institutes, National Institute of Cancer Research, Tainan 70456, Taiwan.
Cells. 2022 Feb 11;11(4):638. doi: 10.3390/cells11040638.
This research aimed to excavate the alternative mechanism of CEBPD on tumor growth and explore the biological significance of the CEBPD/hsa-miR-429/VEGFA axis on angiogenesis in urothelial carcinoma (UC).
Quantitative RT-PCR, immunoblotting assay and tube formation examined the effect of hsa-miR-429 mimic or/and inhibitor on VEGFA expression and angiogenesis in CEBPD-overexpressing UC-derived cells. The association between CEBPD, hsa-miR-429, VEGFA and microvascular density (MVD) and clinical outcome were evaluated in 296 patients with UBUC and 340 patients with UTUC, respectively.
The increase in the transcript and protein of VEGFA as well as HUVECs tube formation was diminished upon the treatment of hsa-miR-429 mimic in CEBPD-overexpressing BFTC909 and TCCSUP. Nevertheless, the inhibited regulation of hsa-miR-429 mimic on the expression of VEGFA and ability of HUVECs tube formation was rescued by the combined incubation with hsa-miR-429 inhibitor in these two UC-derived cell lines. Furthermore, the clinical correlations showed that the higher level of VEGFA or MVD has a positive correlation with the expression of CEBPD and a negative relation to hsa-miR-429 and leads to tumor aggressiveness with worse disease-specific, metastasis-free survival in UBUC and UTUC cohorts.
We decipher the oncogenic mechanism of CEBPD on angiogenesis through the hsa-miR-429 inhibition to stabilize the expression of VEGFA in UC. The novel research unveiled the modulation of the CEBPD/hsa-miR-429/VEGFA axis on the progression of UC and could be accessible to theranostic biomarkers.
本研究旨在挖掘 CEBPD 对肿瘤生长的替代机制，并探讨 CEBPD/hsa-miR-429/VEGFA 轴在尿路上皮癌（UC）血管生成中的生物学意义。
定量 RT-PCR、免疫印迹分析和管形成实验检测了 hsa-miR-429 模拟物或/和抑制剂对 CEBPD 过表达 UC 源性细胞中 VEGFA 表达和血管生成的影响。分别在 296 例膀胱癌（UBUC）和 340 例上尿路上皮癌（UTUC）患者中评估了 CEBPD、hsa-miR-429、VEGFA 和微血管密度（MVD）之间的相关性及其与临床结局的关系。
在 CEBPD 过表达的 BFTC909 和 TCCSUP 细胞中，hsa-miR-429 模拟物处理后，VEGFA 的转录本和蛋白水平以及 HUVEC 管形成均减少。然而，在这两种 UC 源性细胞系中，hsa-miR-429 抑制剂的联合孵育挽救了 hsa-miR-429 模拟物对 VEGFA 表达和 HUVEC 管形成能力的抑制作用。此外，临床相关性研究表明，VEGFA 或 MVD 水平升高与 CEBPD 的表达呈正相关，与 hsa-miR-429 的表达呈负相关，导致 UBUC 和 UTUC 队列肿瘤侵袭性增加，疾病特异性、无转移生存率降低。
我们通过 hsa-miR-429 抑制来稳定 UC 中 VEGFA 的表达，解析了 CEBPD 对血管生成的致癌机制。这项新的研究揭示了 CEBPD/hsa-miR-429/VEGFA 轴对 UC 进展的调控作用，并可作为治疗的生物标志物。
